The effectiveness of clove oil, its fractions, and clove oil-based fumigant tablet formulations, against Tribolium castaneum (Herbst)

Indonesia has seen an increase and widespread reports of resistance among stored-product insect pests to phosphine. The use of Syzigium aromaticum (clove oil) as an alternative fumigant may be a useful strategy to control infestation by phosphine-resistant varieties of stored-product insects. The objective of this study was to examine the effectiveness of whole (unfractionated) clove oil as well as its component fractions as a fumigant and repellent against the red flour beetle (Tribolium castaneum (Herbst)), and to develop a simple fumigant formulation for this purpose. The experimental design used to test the effectiveness of clove oil and its fractionation was a completely randomized design (CRD). Meanwhile, testing the effectiveness of tablet formulations was carried out by factorial CRD. Fumigation test results gave LD50 and LD95 values of 0.234 and 1.142 ml/l respectively, for crude clove oil used in a fumigation chamber against T. castaneum. An n-hexane fraction of clove oil tested under the same conditions was more lethal, causing 95% mortality of T. castaneum at the dose of 0.801 ml/l during fumigation. Finally, tablets containing a set proportion of clove oil and naphthalene (1:1) reached LD91 against T. castaneum after 7 days’ fumigation.

EFFICIENCY OF ORGANIC AND INORGANIC PESTICIDES AGAINST STORED GRAIN INSECT PESTS

The insect pests cause substantial loss of grains in the field and during storage of food grains that worth millions of rupees. Hence, in order to manage stored grain insect pests a combination of organic and inorganic pesticide control materials was selected to check the efficiency against red flour beetle (Tribolium castaneum) and pulse beetle (Callosobruchus chinensis). Total 10 treatments were used, eight different organic bio-pesticides (neem seed, neem leaves, naswar, tobacco, eucalyptus, citrus, dhatura and mint) and one inorganic (naphthalene balls) and control used at 32 ± 2°C and 70 ± 5% RH. The results indicated that three doses of all treatments were used as 5, 10 and 20gm in 50gm grains. Overall, maximum (4.33 ± 1.00%) mortality of red flour beetle was observed using neem seed powder and no (0.0 0 ± 00%) mortality was observed in minte leaves powder. In case of pulse beetle, the maximum (5.07 ± 0.87%) mortality was observed using neem seed powder and minimum (0.66±0.22%) using citrus leaves powder. Maximum (16.26 ± 0.03) antifeedant efficiency of red flour beetle was observed in control group and minimum (6.46±0.65) antifeedant efficiency of red flour beetle was found in dhatura leaves powder at all tested concentrations. Maximum (14.23 ± 0.13) antifeedant efficiency of pulse beetle was determined in control group and and minimum (1.00 ± 0.04) was observed for naphthalene balls at all tested concentrations. Maximum (94.67%) repellency against red flour beetle was observed using neem seed powder and minimum (62.00%) was found in naphathalene balls as compared with their controls. Maximum (81.32%) repellency against pulse beetle was observed using naswar powder and minimum (63.34%) repellency was determined using neem leaves powder as compared with their control group. We concluded that 20% of neem seed powder concentration proved effective to suppress the population of both red flour beetle and pulse beetle.

cDNA Cloning and Partial Characterization of the DJ-1 Gene from Tribolium castaneum

The DJ-1 gene is highly conserved across a wide variety of organisms and it plays a role in anti-oxidative stress mechanisms in cells. The red flour beetle, Tribolium castaneum, is widely used as a model insect species because it is easy to evaluate gene function in this species using RNA interference (RNAi). The T. castaneum DJ-1 (TcDJ-1) sequence is annotated in the T. castaneum genome database; however, the function and characteristics of the TcDJ-1 gene have not been elucidated. Here, we investigated the cDNA sequence of TcDJ-1 and partially characterized its function. First, we examined the TcDJ-1 amino acid sequence and found that it was highly conserved with sequences from other species. TcDJ-1 mRNA expression was higher in the early pupal and adult developmental stages. We evaluated oxidant tolerance in TcDJ-1 knockdown adults using paraquat and found that adults with TcDJ-1 knockdown exhibited increased sensitivity to paraquat. Our findings show that TcDJ-1 has an antioxidant function, as observed for DJ-1 from other insects. Therefore, these results suggest that TcDJ-1 protects against oxidative stress during metamorphosis.

Efficacy of Ozone Gas against All Stages of Red Flour Beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) at Different Temperatures and Exposure Periods

Ozone efficacy (approximately of 600 Mg.hr-1) was evaluated against Red Flour Beetle’s (RFB, Tribolium castium (Herbst)) eggs, larvae, pupae and adults at different temperatures (35, 40 and 45 ° C) and exposure times (1, 2, 4, 6, 8, 10 hours) under laboratory. The control treatments included above thermal levels without ozone at the mentioned exposure periods. The results indicated ozone effectively suppressed hatching with increasing temperature. The hatch egg rates was reduced to 0 % in the treated treatments after 10 h at 45° C. However, mortality rates of RFB were increased with maximizing of the exposure time to ozone. After 10-h exposure to ozone at the lowest temperature 35˚ C, complete mortality (100%) or few survivals of RFB were recorded in the susceptible stages (larvae and adults). Also, the complete mortality of adults and larvae resulted after an exposure to ozone at 40 ° C for 10 hours, compared than the high survival rates at the pest’s stages at 30 and 40° C, even after 10 hours of monitoring time. However the exposure time which is required to 100% mortality was decreased to 4 hours at 45 ° C, comparing to the thermal treatment only (control) which needed more time (6 h) for 100% of mortality. In conclusion, ozone application showed the efficacy on the mortality at all stages of RFB and the temperature was a potential factor enhancing the application of ozone for RFB control.

Efficacy of Entomopathogenic Nematodes Against Three Species of Stored Product Insects

Two specis of entomopathogenic nematodes (EPNs), Steinernema riobravi, symbolized by Sr, and Heterorabditisbacterophora, symbolized by Hb,were tested by using five concentrations: 125, 250, 500, 1000, and 2000 an infectiousphase, ml of the type Sr and four concentrations of 500, 1000, 2000 and 3000infectious phase/mlof the type Hb against Cowpea weevil, Callosobruchus maculates,Khapra beetle, Trogoderma granarium, and Rust red flour beetle Triboliumcastaneum, using three exposure times of 24, 48, and 72 h. The mortalityrate increased withincreasing in concentrations and the exposure period. The results showed that the highest mortality rate was 16.67 whole insects recorded for the 2000 infectious phase/ml with Sr nematodes after 72 hours post- treatment ofC. maculatus. The results showed that the highest rate mortality,which is 20 larvae, was recorded for the infectious phase dose of the genus nematode (Hb), all of which are 500, 1000, 2000 and 3000 gastricphase/ml, and for all exposure periods used between 24 hours and up to 72 hours against the larvae of the T. granarium. The results showed that the highest rate of mortality 10.00 whole insects was recorded for the dose of 3000 Infectious stage/ml and at the exposure time 72 hours, while the lowest rate for this mortality of 1.33 adult insects for rusty flour beetle was recorded for the dose of 500 infectious stage/ml and for the exposure time only 24 hours. The resultsindicated that mortality rate was increasing against any increase in both the exposure time and the dose of nematodes of the genus Hb.